Lysis of bacteria in maxi prep.
vilimf01 at mcrcr6.med.nyu.edu
vilimf01 at mcrcr6.med.nyu.edu
Wed Dec 4 21:46:25 EST 1996
I've seen this problem in the past. In my case, I found that it
wasn't E-coli I was growing up, but som unknown, unlysable organism.
The tipoff is, is the pellet orange-yellowish, or white? If its white
and it doesn't lyse, start again with a new transformation and avoid
the colonies which aren't the right color (this worked for me).
Hope this helps.
Regards SVEN
In article <32A569F2.7802 at infobiogen.fr>, Florence CABON <cabon at infobiogen.fr> writes:
> Dear netters,
>
> I had problems recently with a qiagen maxi prep. I was preparing two
> plasmids. The first prep went OK but I failed to lyse the second one. I
> tried to add some SDS, I added some lysozyme, I warmed the mix. The
> bacteria were still intact in suspension. Two days later we started the
> prep again, draining carefully all the LB medium from the bacterial
> pellet. The result was the same.
> The plasmid is in a classical puc backbone. The bacteria are Xl1blue
> from stratagene
> Any advise ?
>
> ======================================================
> Florence Cabon. CNRS Villejuif, France. cabon at infobiogen.fr
> ======================================================
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